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Activation of MEKK1 by GCK requires the autophosphorylation phosphoacceptor sites T1381 and T1393. Wild-type HA-MEKK1, MEKK1(T1381A), and MEKK1(T1393A) were immunoprecipitated from transfected 293 cells as for Fig. 3B and incubated with purified GST-GCK (Fig. 3A). Control incubations (minus GCK) were performed with GST. GST-SEK1(K129R) was added, and the results were analyzed as for Fig. 3B. The graph shows a phosphorimager quantitation of the results in the autoradiograms.
